Abstract. A retrospective analysis revealed that water quality in the coastal bays of Maryland (USA) has been declining over the past decade, as evidenced by increases in total nitrogenous nutrients and in outbreaks of brown tides caused by the pelagophyte Aureococcus anophagefferns. However, the increases in total nitrogen are not a function of increases in inorganic nitrogen, but rather a function of increases in dissolved organic nitrogen (DON). A near-decadal record (1996)(1997)(1998)(1999)(2000)(2001)(2002)(2003)(2004) demonstrates that an approximate doubling of DON over this time period correlates with a similar increase in total chlorophyll and an even larger increase in the proportion of chlorophyll that is composed of brown tide. Additionally, on an annual basis overall chlorophyll levels and strength of the brown-tide blooms were related to the DON availability that developed during the prior months. Aureococcus anophagefferens is a harmful algal bloom species that preferentially uses DON for its nutrition over inorganic nitrogen forms and thus is a symptom of organic rather than inorganic nitrogen-based eutrophication. These results demonstrate that long-term changes in nutrient quantity and composition have occurred in the coastal bays of Maryland during the past decade and that total phytoplankton biomass, as well as the proliferation of brown tide, are related to these changes. Whether these changes are evidence of a long-term trajectory or represent a shortterm anomaly will be revealed in future monitoring.

Abstract. The mid-Atlantic coastal bays are shallow coastal lagoons, separated from the Atlantic Ocean by barrier sand islands with oceanic exchanges restricted to narrow inlets. The relatively poor flushing of these lagoon systems makes them susceptible to eutrophication resulting from anthropogenic nutrient loadings. An intensive water quality and seagrass monitoring program was initiated to track ecological changes in the Maryland and Virginia coastal bays. The purpose of this study was to analyze existing monitoring data to determine status and trends in eutrophication and to determine any associations between water quality and living resources. Analysis of monitoring program data revealed several trends: (1) decadal decreases in nutrient and chlorophyll concentrations, followed by recently increasing trends; (2) decadal increases in seagrass coverage, followed by a recent period of no change; (3) blooms of macroalgae and brown tide microalgae; and (4) exceedance of water quality thresholds: chlorophyll a (15 lg/L), total nitrogen (0.65 mg/L or 46 lmol/L), total phosphorus (0.037 mg/L or 1.2 lmol/L), and dissolved oxygen (5 mg/L) in many areas within the Maryland coastal bays. The water quality thresholds were based on habitat requirements for living resources (seagrass and fish) and used to calculate a water quality index, which was used to compare the bay segments. Strong gradients in water quality were correlated to changes in seagrass coverage between segments. These factors indicate that these coastal bays are in a state of transition, with a suite of metrics indicating degrading conditions. Continued monitoring and intensified management will be required to avert exacerbation of the observed eutrophication trends. Coastal lagoons worldwide are experiencing similar degrading trends due to increasing human pressures, and assessing status and trends relative to biologically relevant thresholds can assist in determining monitoring and management priorities and goals.

A new immunologically based flow cytometry (IFCM) technique was developed to enumerate Aureococcus anophagefferens, a small pelagophyte alga that is the cause of "brown tides" in bays and estuaries of the mid-Atlantic states along the U.S. coast. The method utilizes a monoclonal antibody conjugated to fluorescein isothiocyanate (FITC-MAb) to label the surface of A. anophagefferens cells which are then detected and enumerated by using a flow cytometer. Optimal conditions for FITC-MAb staining, including solution composition, incubation times, and FITC-MAb concentrations, were determined. The FITC-MAb method was tested for cross-reactivity with nontarget, similarly sized, photoautotrophic protists, and the method was compared to an enzyme-linked immunosorbent assay (ELISA) using the same MAb. Comparisons of the IFCM technique to traditional microscopy enumeration of cultures and spiked environmental samples showed consistent agreement over several orders of magnitude (r 2 > 0.99). Comparisons of the IFCM and ELISA techniques for enumerating cells from a predation experiment showed a substantial overestimation (up to 10 times higher) of the ELISA in the presence of consumers of A. anophagefferens, presumably due to egested cell fragments that retained antigenicity, using the ELISA method, but were not characterized as whole algal cells by the IFCM method. Application of the IFCM method to environmental "brown-tide" samples taken from the coastal bays of Maryland demonstrated its efficacy in resolving A. anophagefferens abundance levels throughout the course of a bloom and over a large range of abundance values. IFCM counts of the brown-tide alga from natural samples were consistently lower than those obtained using the ELISA method and were equivalent to those of the polyclonal immunofluorescence microscopy technique, since both methods discriminate intact cells. Overall, the IFCM approach was an accurate and relatively simple technique for the rapid enumeration of A. anophagefferens in natural samples over a wide range of abundance values (10 3 to 10 6 cells ml ؊1 ).

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